Exponential-phase expression of spvA of the Salmonella typhimurium virulence plasmid:: induction in intracellular salts medium and intracellularly in mice and cultured mammalian cells

被引:34
作者
Wilson, JA [1 ]
Doyle, TJ [1 ]
Gulig, PA [1 ]
机构
[1] Univ Florida, Coll Med, Dept Mol Genet & Microbiol, Gainesville, FL 32610 USA
来源
MICROBIOLOGY-UK | 1997年 / 143卷
关键词
Salmonella typhimurium; virulence; regulation; expression;
D O I
10.1099/00221287-143-12-3827
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
The spy genes of Salmonella typhimurium and other non-typhoidal Salmonella serovars are essential for efficient systemic infection beyond the intestines in orally inoculated mice as a model for enteric fever. These virulence genes are not significantly expressed by salmonellae during exponential growth in L broth but are induced when the bacteria enter the stationary phase of growth. Using RNase protection analysis to directly measure spvA mRNA from the virulence plasmid of S. typhimurium, we found that spvA was maximally induced in an SpvR- and RpoS-dependent manner during exponential growth in Intracellular Salts Medium, which mimics the intracellular environment of mammalian cells. A cloned spvA-lacZ operon fusion in S. typhimurium was induced intracellularly in peritoneal cells of mice, correlating in vivo intracellular gene expression with intracellular function of the spy genes in infected mice. spvA was also induced intracellularly in vitro within both Henle-407 intestinal epithelial cells and J774.A1 macrophage-like cells when the bacteria were replicating with exponential kinetics. Prevention of invasion of salmonellae with cytochalasin D inhibited spvA induction within tissue culture cells, indicating that salmonellae must be internalized for spvA to be induced. The spvA-lacZ fusion was not induced by salmonellae in extracellular fluid of the peritoneal cavity or in serum. Since induction of the spy genes occurs intracellularly during exponential growth of salmonellae. cessation of growth may not be the most relevant inducing signal for spy gene expression.
引用
收藏
页码:3827 / 3839
页数:13
相关论文
共 62 条
[1]   REGULATION OF SPVR GENE-EXPRESSION OF SALMONELLA VIRULENCE PLASMID PKDSC50 IN SALMONELLA-CHOLERAESUIS SEROVAR CHOLERAESUIS [J].
ABE, A ;
MATSUI, H ;
DANBARA, H ;
TANAKA, K ;
TAKAHASHI, H ;
KAWAHARA, K .
MOLECULAR MICROBIOLOGY, 1994, 12 (05) :779-787
[2]   GROWTH-RATE PARADOX OF SALMONELLA-TYPHIMURIUM WITHIN HOST MACROPHAGES [J].
ABSHIRE, KZ ;
NEIDHARDT, FC .
JOURNAL OF BACTERIOLOGY, 1993, 175 (12) :3744-3748
[3]  
Ausubel FM., 1993, Current Protocols in Molecular Biology
[4]   STATIONARY-PHASE-INDUCIBLE GEARBOX PROMOTERS - DIFFERENTIAL-EFFECTS OF KATF MUTATIONS AND ROLE OF SIGMA-70 [J].
BOHANNON, DE ;
CONNELL, N ;
KEENER, J ;
TORMO, A ;
ESPINOSAURGEL, M ;
ZAMBRANO, MM ;
KOLTER, R .
JOURNAL OF BACTERIOLOGY, 1991, 173 (14) :4482-4492
[5]   THE SALMONELLA-TYPHIMURIUM VIRULENCE PLASMID ENCODES A POSITIVE REGULATOR OF A PLASMID-ENCODED VIRULENCE GENE [J].
CALDWELL, AL ;
GULIG, PA .
JOURNAL OF BACTERIOLOGY, 1991, 173 (22) :7176-7185
[6]   Expression of Salmonella typhimurium rpoS and rpoS-dependent genes in the intracellular environment of eukaryotic cells [J].
Chen, CY ;
Eckmann, L ;
Libby, SJ ;
Fang, FC ;
Okamoto, S ;
Kagnoff, MF ;
Fierer, J ;
Guiney, DG .
INFECTION AND IMMUNITY, 1996, 64 (11) :4739-4743
[7]   CENTRAL REGULATORY ROLE FOR THE RPOS SIGMA-FACTOR IN EXPRESSION OF SALMONELLA-DUBLIN PLASMID VIRULENCE GENES [J].
CHEN, CY ;
BUCHMEIER, NA ;
LIBBY, S ;
FANG, FC ;
KRAUSE, M ;
GUINEY, DG .
JOURNAL OF BACTERIOLOGY, 1995, 177 (18) :5303-5309
[8]   Salmonella spp are cytotoxic for cultured macrophages [J].
Chen, LM ;
Kaniga, K ;
Galan, JE .
MOLECULAR MICROBIOLOGY, 1996, 21 (05) :1101-1115
[9]   GROWTH-PHASE AND SPVR REGULATION OF TRANSCRIPTION OF SALMONELLA-TYPHIMURIUM SPVABC VIRULENCE GENES [J].
COYNAULT, C ;
ROBBESAULE, V ;
POPOFF, MY ;
NOREL, F .
MICROBIAL PATHOGENESIS, 1992, 13 (02) :133-143
[10]   GENE ALGD CODING FOR GDPMANNOSE DEHYDROGENASE IS TRANSCRIPTIONALLY ACTIVATED IN MUCOID PSEUDOMONAS-AERUGINOSA [J].
DERETIC, V ;
GILL, JF ;
CHAKRABARTY, AM .
JOURNAL OF BACTERIOLOGY, 1987, 169 (01) :351-358